一种含特异性侵染内共生菌Rickettisa massilia 的烟粉虱种群建立方法
A method for establishing a Bemisia tabaci population infected with Rickettisa massilia
杨义婷** 张 焱 郭建洋 郭建英*** 万方浩***
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DOI：10.7679/j.issn.2095-1353.2015.029
作者单位：中国农业科学院植物保护研究所，植物病虫害生物学国家重点实验室，北京 100193
中文关键词：烟粉虱，内共生菌，Rickettisa massilia，种群建立
英文关键词： Bemisia tabaci, endosymbionts, Rickettisa, population establishment
中文摘要：【目的】 烟粉虱Bemisia
tabaci（Gennadaius）广泛分布于全球除南极洲外各大洲的90多个国家和地区，其寄主范围广，危害严重，属于农业上的重要害虫。烟粉虱体内的内共生菌有助于其扩散传播，提高其寄主适合度，但其体内的内共生菌大都为次生内共生菌，很难进行体外培养，因此，加大了其内共生菌研究的难度。【方法】 采集我国部分地区的烟粉虱种群，然后利用基于mtDNA COⅠ基因的种特异性引物进行种群生物型鉴定，再用内共生菌的特异性引物来鉴定含有特异性侵染内共生菌种群，确定个体含内生菌类型后，采取继代饲养方法建立种群。【结果】 经鉴定烟粉虱的生物型为Q型（MED隐种），经过5代种群筛选后，可以建立稳定遗传的含特异性侵染内共生菌Rickettisa
massilia菌的烟粉虱种群。【结论】 通过本方法可以获得稳定的含有特异性侵染内共生菌Rickettisa
massilia菌的烟粉虱种群。该方法对研究烟粉虱与体内内共生菌、植物三者之间以及烟粉虱体内某几种内共生菌之间的互作有重要意义，同时也为明确烟粉虱体内Rickettisa菌的作用研究打下基础。
英文摘要：[Objectives] The whitefly, Bemisia tabaci (Gennadaius), is widely distributed in over more than 90 countries and all the world’s regions except Antarctica. Because of its wide host range and capacity to cause serious damage to agricultural crops this species is a particularly destructive pest. The endosymbionts of B. tabaci contribute both to its distribution and host fitness. However, most of these endosymbionts are secondary ones that are very difficult to culture in vitro. [Methods] Whiteflies were collected and reared in a laboratory. One pair of species-specific PCR primers based on a fragment of the known mitochondrial DNA cytochrome oxidaseⅠ(mtDNA COⅠ)sequence was used to diagnose the biotype of the collected specimens. Next, a specific primer was designed to detect whitefly endosymbionts. [Results] The biotype of the B. tabaci specimens collected was MED (previously called the Q biotype). After five generations of population screening, a stable whitefly population carrying and transmitting the endosymbiont Rickettsia to subsequent generations was established. [Conclusion] The method can produce a stable Rickettsia-infected whitefly population, which is useful for studying the interactions between whiteflies, their endosymbionts, and host plants. This method lays a foundation for investigating the role of Rickettsia in the biology of B. tabaci.